We have undertaken the following projects: Autoimmune diseases: subclass specific characterization of IgG glycosylation in myositis. Aims: 1) To determine whether a pattern of selected agalactosyl IgG glycoforms is characteristic of myositis patients similar to that found in rheumatoid arthritis patients. 2) Determine if IgG markers of susceptibility can be identified. Results: 1) Elevated amounts of glycoforms lacking terminal galactose, IgG-Gal 0, were found in myositis patients. 2 )Statistical trend analysis indicates an increasing pattern in IgG-Gal 0 in the order controls &#8804;asymptomatic siblings &#8804;patients in both IgG1 and IgG2-3. Characterizatioon of O-glycosylation in collagen Aims: 1) identification of sites of O-linked glycosylation in type I collagen from mouse osteoblast cells and bovine bone;2) structural characterization of bi- (DHLNL) and trifunctional (Pyr) cross-linked peptides in human and bovine collagen, including their glycosylation status. 3) characterization of glycosylation changes in mouse type I collagen from LH3 suppressed osteoblast clones. Results: ) site specific occupancies were identified. 2) five cross-linked species (3 divalent, 2 trivalent) were found in bovine bone collagen. 2 trivalent species found in human bone collagen. Glycosylation states were identified. 3) Glycosylation in LH3 suppressed osteoblasts were observed to be significantly lower than in wt. Anthrax studies Aims: To determine whether or not the spin-trapping agent, DMPO, can alter the course of anthrax lethal toxin intoxication. Results: Based on previous studies that indicate that free-radical induced oxidation is a major player in anthrax, we studied the effect on LT-exposed macrophage cells of DMPO dosing at several levels and time-points post exposure to LT. We found that cell viability increased significantly at DMPO doses up to 100 mM and up to 1 hr dosing post exposure. Further experiments found that this effect was dependent on the source/purity of DMPO. We are currently trying to identify the component responsible for the protective effect.